Purification by affinity chromatography and physicochemical properties of the guanine-specific ribonuclease of Fusarium moniliforme.
Ribonuclease F1, the guanine-specific ribonuclease of Fusarium moniliforme, was purified to homogeneity by a combination of ethanol fractionation, affinity chromatography and DEAE-cellulose column chromatography. The adsorbent for the affinity chromatography was synthesized by the coupling of periodate-oxidized guanosine 5'-monophosphate to aminohexyl agarose followed by sodium borohydride reduction. Ribonuclease F2, the minor component, was also purified to near homogeneity by the same procedure. Ribonucleases F1 and F2 had the same molecular weight (about 11,000) as determined by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. They also showed the same amino acid composition and differed only in the isoelectric point: 4.10 for F1 and 3.96 for F2.
D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D005670:Fusarium / Q000201:enzymology*; D007526:Isoelectric Point; D008970:Molecular Weight; D006163:Ribonuclease T1 / Q000302:isolation & purification*; D012260:Ribonucleases / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D012260:Ribonucleases; D006163:Ribonuclease T1